Etiologic Risk Group (see link below): Appendix B-III-A. Risk Group 3 (RG3) - Bacterial Agents Including Rickettsia
*/Disease Information (sort of like the Intro to your Mini Research Write up): The disease Tuberculosis is caused by Mycobacterium tuberculosis. The bacteria usually affects the lungs, but can also affect other parts of the body such as the kidneys. Tuberculosis may be fatal if not treated. Tuberculosis is spread through the air, making it very easy to contract. Symptoms could arise within 2-12 weeks of exposure or may not be seen until a year; this is known as latent TB infection. Symptoms include coughing up blood, chest pain, weight loss, chills, fever, etc. HIV infections increase the likelihood of the latent TB to show symptoms. One-third of the world's population is thought to have been infected with M. tuberculosis, and new infections occur in about 1% of the population each year.
Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): Coupled Enzyme assay AGK activity was determined by a coupled assay with AGPR measuring the decrease of NADPH by its absorbance at 340 nm. The reaction mixture contained 100 mM potassium phosphate buffer (pH 7.0), 10 mM N-acetylglutamate, 10 mM ATP, 20 mM MgCl2, 0.2 mM NADPH, and 10 μg AGPR. The reaction was started by the addition of N-acetylglutamate. Unless stated otherwise, a reaction mixture without N-acetylglutamate served as a reference in the spectrophotometric assay. One unit of AGK activity corresponded to the oxidation of 1 μmol of NADPH per minute at 30 °C. http://www.sciencedirect.com/science/article/pii/S0022283607000733 --- List cost and quantity of substrate reagents, supplier, and catalog #
Structure (PDB or Homology model) -- PDB #: 2I3G
Current Inhibitors:
Expression Information (has it been expressed in bacterial cells): Expressed in E. coli cells
Purification Method: Hanging drop vapor diffusion technique
Length of protein in Amino Acids: 352 Molecular Weight of protein: 2 chains- 72.5906 kDa, 1 chain- 36.3043 kDa Molar Extinction coefficient of your protein at 280 nm wavelength: 2 chains- 60195 1 chain- 30035 TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html).
*GC% Content for gene: 69.7% *CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only): *GC% Content for gene (codon optimized):
*NCBI Gene # or RefSeq#: 885278
*Protein ID (NP or XP #) or Wolbachia#: NP_216168.1
*Organism (including strain): Mycobacterium tuberculosis
Etiologic Risk Group (see link below): Appendix B-III-A. Risk Group 3 (RG3) - Bacterial Agents Including Rickettsia
*/Disease Information (sort of like the Intro to your Mini Research Write up):
The disease Tuberculosis is caused by Mycobacterium tuberculosis. The bacteria usually affects the lungs, but can also affect other parts of the body such as the kidneys. Tuberculosis may be fatal if not treated. Tuberculosis is spread through the air, making it very easy to contract. Symptoms could arise within 2-12 weeks of exposure or may not be seen until a year; this is known as latent TB infection. Symptoms include coughing up blood, chest pain, weight loss, chills, fever, etc. HIV infections increase the likelihood of the latent TB to show symptoms. One-third of the world's population is thought to have been infected with M. tuberculosis, and new infections occur in about 1% of the population each year.
Link to TDR Targets page (if present): N/A
Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.):
http://www.ncbi.nlm.nih.gov/pubmed/17316682?dopt=Abstract
Essentiality of this protein: Essential for optimal growth of the Mycobacterium tuberculosis pathogen
http://www.sciencedirect.com/science/article/pii/S0022283607000733
Complex of proteins?: homotetramer
Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism):
http://www.sciencedirect.com/science/article/pii/S0022283607000733
*EC#:1.2.1.38
Link to BRENDA EC# page:
http://www.brenda-enzymes.org/enzyme.php?ecno=1.2.1.38&Suchword=&organism%5B%5D=Mycobacterium+tuberculosis&show_tm=0
Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): Coupled Enzyme assay
AGK activity was determined by a coupled assay with AGPR measuring the decrease of NADPH by its absorbance at 340 nm. The reaction mixture contained 100 mM potassium phosphate buffer (pH 7.0), 10 mM N-acetylglutamate, 10 mM ATP, 20 mM MgCl2, 0.2 mM NADPH, and 10 μg AGPR. The reaction was started by the addition of N-acetylglutamate. Unless stated otherwise, a reaction mixture without N-acetylglutamate served as a reference in the spectrophotometric assay. One unit of AGK activity corresponded to the oxidation of 1 μmol of NADPH per minute at 30 °C.
http://www.sciencedirect.com/science/article/pii/S0022283607000733
--- List cost and quantity of substrate reagents, supplier, and catalog #
Structure (PDB or Homology model)
-- PDB #: 2I3G
Current Inhibitors:
Expression Information (has it been expressed in bacterial cells): Expressed in E. coli cells
Purification Method: Hanging drop vapor diffusion technique
http://scripts.iucr.org/cgi-bin/paper?S0907444902020802
Image of protein (PyMol with features delineated and shown separately):
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):
Length of protein in Amino Acids: 352
Molecular Weight of protein: 2 chains- 72.5906 kDa, 1 chain- 36.3043 kDa
Molar Extinction coefficient of your protein at 280 nm wavelength: 2 chains- 60195 1 chain- 30035
TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html).
*CDS Gene Sequence (paste as text only):
*GC% Content for gene: 69.7%
*CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only):
*GC% Content for gene (codon optimized):